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Plasmid Files

pIEx™-2

Vector for high-level expression in insect cells of proteins with a cleavable N-terminal GST-6xHis-S-Tag cassette plus a C-terminal HSV tag.

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pIEx-2.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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DrdI (4095) PspFI (3897) BseYI (3893) AhdI (3309) BsaI (3243) BpmI (3240) NmeAIII (3162) AatII (2390) ZraI (2388) XbaI (2382) PflFI - Tth111I (2327) BspDI * - ClaI * (207) EcoRV (239) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BsiWI (784) MluI (872) BtgI - NcoI (1082) EcoNI (1097) EcoO109I (1119) MscI (1294) BsgI (1373) SwaI (1514) SpeI (1753) XcmI (1786) PflMI (1830) PshAI (1904) BseRI (1908) BamHI (1926) MfeI (1932) Eco53kI (1941) BanII - SacI (1943) BsrGI (1946) AscI (1954) PstI - SbfI (1964) SalI (1966) Acc65I (1972) AgeI (1975) KpnI (1976) HindIII (1984) NotI (1991) BstZ17I (2006) PmlI (2011) BbsI (2045) PaeR7I - XhoI (2053) DraIII (2064) Bsu36I (2074) PacI (2103) pIEx™-2 4563 bp
DrdI  (4095)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PspFI  (3897)
1 site
C C C A G C G G G T C G
BseYI  (3893)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AhdI  (3309)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BsaI  (3243)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BpmI  (3240)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
NmeAIII  (3162)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AatII  (2390)
1 site
G A C G T C C T G C A G
ZraI  (2388)
1 site
G A C G T C C T G C A G
XbaI  (2382)
1 site
T C T A G A A G A T C T
PflFI  (2327)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2327)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BspDI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoRV  (239)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
PsiI  (409)
1 site
T T A T A A A A T A T T
NheI  (606)
1 site
G C T A G C C G A T C G
BmtI  (610)
1 site
G C T A G C C G A T C G
SphI  (614)
1 site
G C A T G C C G T A C G
BsmI  (775)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
BsiWI  (784)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
MluI  (872)
1 site
A C G C G T T G C G C A
BtgI  (1082)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1082)
1 site
C C A T G G G G T A C C
EcoNI  (1097)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
EcoO109I  (1119)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
MscI  (1294)
1 site
T G G C C A